Compositions and methods for the sustained release of betaalanine

ABSTRACT

Methods and compositions are provided for increasing the anaerobic working capacity of muscle and other tissues. Also provided are compositions formulated for the sustained release of free beta-alanine. Also provided is a dietary supplement formulated, for example, as a solid food product, an edible suspension, liquid or semi-liquid as described herein.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.14/626,739, filed Feb. 19, 2015, which is a continuation of U.S.application Ser. No. 13/929,251, filed Jun. 27, 2013, now U.S. Pat. No.8,980,307, issued Mar. 17, 2015, which is a continuation of U.S.application Ser. No. 13/755,954, filed Jan. 31, 2013, now U.S. Pat. No.8,496,958, issued Jul. 30, 2013, which is a continuation of U.S.application Ser. No. 13/646,184, filed Oct. 5, 2012, now U.S. Pat. No.8,394,402, issued Mar. 12, 2013, which is a divisional of U.S.application Ser. No. 12/064,005, filed Apr. 30, 2009, now U.S. Pat. No.8,329,207, issued Dec. 11, 2012, which is a National Stage applicationunder 35 U.S.C. § 371, which claims benefit under 35 U.S.C. § 119(a) ofInternational Application No. PCT/US2006/020104, filed May 23, 2006,which claims benefit of U.S. Provisional Patent Application No.60/683,985, filed May 23, 2005, all of which are herein incorporated byreference.

TECHNICAL FIELD

This application relates generally to the fields of pharmaceuticals andphysiology and more specifically to compositions and methods for thesustained release of free beta-alanine.

BACKGROUND

Natural food supplements are typically designed to compensate forreduced levels of nutrients in the modern human and animal diet. Inparticular, useful supplements increase the function of tissues whenconsumed. It can be particularly important to supplement the diets ofhumans and particular classes of animals whose normal diet may bedeficient in nutrients available only from meat and animal products(e.g. human vegetarians and other animals consuming an herbivorousdiet).

In addition, in the sporting and athletic community, natural foodsupplements which specifically improve athletic ability are increasinglyimportant, such as supplements that promote or enhance physical prowessfor leisure or employment purposes. In another example, anaerobic (e.g.,lactate producing) stress can cause the onset of fatigue and discomfortthat can be experienced with aging. Anaerobic stress can also resultfrom prolonged sub-maximal isometric exercise when the local circulationis partially or totally occluded by the increase in intra-muscularpressure (e.g., during rock climbing) or from exercise involvingprolonged breath holding (e.g., free diving, or synchronized swimming).Excessive lactate production can result in the acidification of theintracellular environment.

Previous work demonstrated that carnosine is significantly increased inmuscle following administration of beta-alanine or a biological sourceof beta-alanine (e.g., carnosine) supplementation. The dosing schedulethat has been preferred for such administration (i.e., 8 times daily),however, leads to compliance issues and problems with paraesthesia.Thus, there is a need for improved supplements containing freebeta-alanine or a biological source thereof in a sustained releaseformulation to increase compliance and decrease paraesthesia events.

SUMMARY

Administration of a bolus single dose of 2-6 grams of free beta-alanine(or chemically equivalent amounts of compounds which are biologicalsources of beta-alanine, e.g., carnosine) results in paraesthesia, asignificant adverse side-effect with symptoms that include abnormalsensations such as burning, tingling, or a “pins-and-needles” feeling.Sustained release of free beta-alanine directly or from compounds whichcan act as biological sources of beta-alanine would obviate seriousadverse side-effects by attenuating the rise in the blood-plasmaconcentration of beta-alanine, providing a means to safely andeffectively increase carnosine biosynthesis in muscle and to attenuatemetabolic acidosis and muscle fatigue during anaerobic activity.

Provided herein are compositions and methods for the sustained releaseof free beta-alanine or a biological source thereof. In one embodiment,the composition is a pharmaceutical composition containing freebeta-alanine or a biological source thereof and an acceptable carrier,where the composition is formulated for the sustained release of freebeta-alanine. In another embodiment, the composition is a dietarysupplement containing free beta-alanine or a biological source thereof.

Also provided are methods for increasing the anaerobic working capacityof muscle and other tissues. Such methods include providing acomposition formulated for the sustained release of free beta-alanine,and administering free beta-alanine or a biological source thereof in aneffective amount to increase the synthesis of carnosine in muscle orother tissues.

Pharmaceutical compositions containing free beta-alanine or a biologicalsource thereof and a pharmaceutically acceptable carrier formulated forsustained release of free beta-alanine are provided. In one embodiment,the pharmaceutical composition is a tablet, capsule, or microbeads thatcan be administered orally. In another embodiment, the pharmaceuticalcomposition is a skin patch or a topical dermatological compoundformulated for the sustained release of free beta-alanine. Thepharmaceutical composition can be formulated for humans.

Dietary supplements containing free beta-alanine or a biological sourcethereof are provided. In one embodiment, the dietary supplement is anenergy bar, edible suspension or other food product. The dietarysupplement can be formulated for humans.

The pharmaceutical composition can include about 0.1-200.0; about1.0-9.0; about 2.0-8.0; about 3.0-7.0; or about 4.0-6.0 grams (g) offree beta-alanine or the equivalent amount of a biological sourcethereof. The pharmaceutical composition can be formulated for humans andcan include about 0.1-200.0; about 1.0-9.0; about 2.0-8.0; about3.0-7.0; or about 4.0-6.0 g of free beta-alanine or the equivalentamount of a biological source thereof.

A dietary supplement can include about 0.1-200.0; about 1.0-9.0; about2.0-8.0; about 3.0-7.0; or about 4.0-6.0 g of free beta-alanine or theequivalent amount of a biological source thereof. The dietary supplementcan be formulated for humans and can include about 0.1-200.0; about1.0-9.0; about 2.0-8.0; about 3.0-7.0; or about 4.0-6.0 g of freebeta-alanine or the equivalent amount of a biological source thereof. Inone embodiment, the dietary supplement is an energy bar, ediblesuspension or food stuff. In one aspect, the dietary supplement containsmultiple servings, and containing 0.1-10.0 g of beta-alanine per servingor the equivalent amount of a biological source thereof. The totaldosage of free beta-alanine or the equivalent amount of a biologicalsource thereof contained in the dietary supplement can be about0.1-200.0; about 1.0-8.0; about 3.0-7.0; or about 4.0-6.0 g of freebeta-alanine or the equivalent amount of a biological source thereof.

Also provided herein are formulations for the sustained release of freebeta-alanine or an equivalent amount of a biological source thereof thatare contained in pharmaceutical compositions. In one aspect, apharmaceutical composition containing free beta-alanine or an equivalentamount of a biological source thereof is formulated for sustainedrelease and can be administered orally in the form of a tablet, gel, ormicrobeads. In another aspect, the pharmaceutical composition is atopical dermatological formulation such as, for example, a hydrogelformulated for the sustained release of free beta-alanine across theskin.

While not limited by any particular mechanism of action, provided hereinare methods and compositions for increasing the anaerobic workingcapacity of muscle and other tissues. The methods and compositionsdisclosed herein provide for the accumulation of carnosine within atissue in the body, resulting from the availability of beta-alanine. Themethods include ingesting or infusing compositions containing freebeta-alanine or a biological source thereof formulated for sustainedrelease into the body. In one aspect, the compositions are mixtures ofcompounds capable of increasing the accumulation of beta-alanylhistidinepeptides (e.g., carnosine) in human and animal tissue. The compositionscan induce the synthesis and accumulation of beta-alanylhistidinepeptides in a human or animal body when introduced into the body.

The compositions provided herein can be used for the preparation of adietary supplement including, e.g., drinks, edible suspensions or foods,or pharmaceutical composition for humans or animals. Compositions of theinvention can be used in any of the methods of the invention.

In one aspect, methods are provided for increasing the anaerobic workingcapacity of a tissue. The method includes the steps of providing freebeta-alanine to blood or blood plasma in an amount effective to increasecarnosine synthesis in a tissue. The tissue referred to herein can beskeletal muscle.

Therefore, in an exemplary aspect, provided herein are compositions andmethods for the sustained release of free beta-alanine comprised in apharmaceutical composition or in a dietary supplement to increase theanaerobic working capacity of muscle tissue as a result of increasedcarnosine biosynthesis.

Unless otherwise defined, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention belongs. Although methods and materialssimilar or equivalent to those described herein can be used in thepractice or testing of the present invention, suitable methods andmaterials are described below. In addition, the materials, methods, andexamples are illustrative only and not intended to be limiting. Allpublications, patent applications, patents, and other referencesmentioned herein are incorporated by reference in their entirety. Incase of conflict, the present specification, including definitions, willcontrol.

The details of one or more embodiments of the invention are set forth inthe accompanying drawings and the description below. Other features,objects, and advantages of the invention will be apparent from thedrawings and detailed description, and from the claims.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a graph showing the blood plasma levels of beta-alanine inindividuals after ingesting either a beta-alanine drink or beta-alaninesustained release capsules.

FIG. 2 is a graph showing the blood plasma levels of beta-alanine inindividuals following ingestion of two different sustained releasebeta-alanine formulations.

Like reference symbols in the various drawings indicate like elements.

DETAILED DESCRIPTION

Provided herein are compositions for the sustained release of freebeta-alanine. Also provided herein are methods for increasing theanaerobic working capacity of a tissue by providing free beta-alanine toblood or blood plasma in an amount that is effective to increase thesynthesis of beta-alanylhistidine peptides, particularly carnosine, in atissue. Beta-alanylhistidine peptides can include peptides ofbeta-alanine, such as carnosine, anserine, and balenine. Sustainedrelease of free beta-alanine is desirable to prevent unwanted adverseside effects such as paraesthesia.

Any of the beta-alanine-containing compositions disclosed herein can beformulated as the sole pharmaceutically active ingredient in thecomposition, or beta-alanine may be combined and/or co-administered withother active ingredients. For example, another active ingredient may beanother pharmacological agent known in the general art to be of value toattenuate the metabolic acidosis encountered during high-intensityexercise by increasing carnosine biosynthesis to delay the onset offatigue during anaerobic exercise. In addition, combinations withcreatine are desirable because the ergogenic effect of the formulationis enhanced. Further, the active materials also can be mixed with otheractive materials which do not impair the desired action, or withmaterials that supplement the desired action, such as antacids, H2blockers, and diuretics.

Co-administration as used herein refers to the administration of morethan one therapeutic agents in the course of a coordinated treatment toachieve an improved clinical outcome. Such co-administration may also becoextensive; that is, occurring during overlapping periods of time.

A. Beta-Alanine and Muscles

Beta-alanine and L-histidine and their methylated analogues formdi-peptides within the human or animal body. The di-peptides producedfrom beta-alanine and histidine include carnosine(beta-alanyl-L-histidine), anserine (beta-alanyl-L-1-methylhistidine),or balenine (beta-alanyl-L-3-methylhistidine) (referred to genericallyherein as “beta-alanylhistidine peptides”). Beta-alanylhistidinepeptides are involved in the regulation of intra-cellular pH homeostasisduring muscle contraction and, therefore, are involved in thedevelopment of muscle fatigue. Beta-alanylhistidine peptides provide aneffective way of accumulating pH-sensitive histidine residues in a cell.Thus, variations in the muscle beta-alanylhistidine peptideconcentrations affect the anaerobic work capacity of muscles, andincreasing the amount of beta-alanylhistidine peptides within a musclefavorably affects performance and the amount of work that can beperformed by the muscle.

Beta-alanine and L-histidine can be generated within the body or aremade available via the diet. Within the body, beta-alanine istransported to tissues such as muscle. Since in a typical fed state, theconcentration of beta-alanine in muscle is low in comparison with theconcentration of L-histidine, the concentration of beta-alanine islikely limiting to the synthesis of beta-alanylhistidine peptides. Thesynthesis and accumulation of beta-alanylhistidine peptides in a humanor animal body can be increased by increasing the blood or blood plasmaconcentrations of beta-alanine, increasing the blood or blood plasmaconcentrations of beta-alanine and creatine, or increasing the blood orblood plasma concentrations of beta-alanine, L-histidine, and creatine(i.e., N-(aminoiminomethyl)-N-glycine, N-amidinosarcosine,N-methyl-N-guanylglycine, or methylglycocyamine).

During sustained intensive exercise or exercise sustained underconditions of local hypoxia, the accumulation of hydronium ions formedduring glycolysis and the accumulation of lactate due to anaerobicmetabolism can severely reduce the intracellular pH. The reduced pH cancompromise the function of the creatine-phosphorylcreatine system. Thedecline in intracellular pH also can affect other functions within thecells such as the function of the contractile proteins in muscle fibers.Administering beta-alanine to an individual elevated musclebeta-alanylhistidine peptide levels and increased total work capacity ofthe muscle. In addition, chronic dietary supplementation withbeta-alanine can increase muscle beta-alanylhistidine peptideconcentration and, thus, increase intramuscular buffering capacity.

Providing beta-alanine or a compound that is a biological source ofbeta-alanine (e.g., carnosine, anserine, or balanine, and salts andchemical derivatives thereof) is a feasible means by which to attenuatethe metabolic acidosis encountered during high-intensity exercise byincreasing beta-alanylhistidine peptide biosynthesis and, thus, delaythe onset of fatigue during anaerobic exercise. A compound that is abiological source of beta-alanine is defined herein as a compound that,when administered to the body by any route (for example parenterally,orally, topically), is converted by one or more chemical- orenzyme-catalysed reaction steps to beta-alanine, which then appears inblood, plasma or serum and is available for uptake into muscle and othertissues.

The free beta-alanine content of the composition may be present as anyactive form of beta-alanine or analogues thereof (e.g., esters orsalts). The free beta-alanine content of the composition can besubjected to a micronization process (e.g. crushing, pulverizing,powdering and the like) prior to incorporation into the pharmaceuticalcomposition so that the resulting composition is not unacceptably“gritty” in texture.

B. Pharmaceutical Compositions

The compositions provided herein include free beta-alanine or abiological source thereof in combination with a pharmaceuticallyacceptable carrier that is formulated for sustained release. While onemode of delivery is oral, other modes of delivery of compositionsaccording to this invention can be used. These include mucosal delivery,nasal delivery, ocular delivery, transdermal delivery, parenteraldelivery, vaginal delivery, rectal delivery, and intrauterine delivery.The compositions can be in a liquid, semi-liquid or solid form, and canbe formulated in a manner suitable for the intended route ofadministration.

Typically, compositions containing free beta-alanine or a biologicalsource thereof as described herein can be formulated into pharmaceuticalcompositions using techniques and procedures well known in the art. See,e.g., Ansel, Introduction to Pharmaceutical Dosage Forms, 4^(th) Ed.,1985, pg 126. Pharmaceutical carriers or vehicles suitable foradministration of free beta-alanine or a biological source thereofinclude any such carriers known to those skilled in the art that aresuitable for the particular mode of administration.

Formulations containing suitable quantities of beta-alanine or abiological source thereof include, for example, capsules, tablets,pills, powders, granules, sterile parenteral solutions or suspensions,oral solutions or suspensions, oil water emulsions as well as implantsand microencapsulated delivery systems, and biodegradable, biocompatiblepolymers such as collagen, ethylene vinyl acetate, polyanhydrides,polyglycolic acid, polyorthoesters, and polylactic acid. Methods forpreparing these compositions are known to those skilled in the art.

Formulations for oral delivery can include, for example, tablets,capsules, granules, and bulk powders. Types of oral tablets includecompressed, chewable lozenges and tablets which may be enteric coated,sugar coated or film coated. Capsules can be hard or soft gelatincapsules, while granules and powders can be provided in non-effervescentor effervescent form with a combination of other ingredients known tothose skilled in the art. By way of example, free beta-alanine or abiological source thereof can be suspended in, for example, propylenecarbonate, vegetable oils or triglycerides and encapsulated in a gelatincapsule. Such solutions and the preparation and encapsulation thereofare disclosed, for example, in U.S. Pat. Nos. 4,328,245; 4,409,239; and4,410,545.

By way of example, enteric coated tablets, because of the entericcoating, resist the action of stomach acid and dissolve or disintegratein the neutral or alkaline intestines; sugar coated tablets arecompressed tablets to which different layers of pharmaceuticallyacceptable substances are applied; film coated tablets are compressedtablets which have been coated with a polymer or other suitable coating;and multiple compressed tablets are compressed tablets made by more thanone compression cycle utilizing the pharmaceutically acceptablesubstances previously mentioned. Any of these formulations of tabletsare suitable for delivering beta-alanine or a biological source thereof.

Free beta-alanine or a biological source thereof may be orallyadministered in a liquid form. Liquid formulations for oral deliveryinclude aqueous solutions (e.g., elixirs and syrups), emulsions,suspensions, solutions and/or suspensions. Elixirs are clear, sweetened,hydroalcoholic preparations, and pharmaceutically acceptable carriersused in elixirs generally include solvents. Syrups are concentratedaqueous solutions of a sugar, for example, sucrose, and may contain apreservative. An emulsion is a two phase system in which one liquid isdispersed in the form of small globules throughout another liquid.Pharmaceutically acceptable carriers used in emulsions are non aqueousliquids, emulsifying agents and preservatives. Suspensions usepharmaceutically acceptable suspending agents and preservatives.

For the preparation of solutions or suspensions, it is possible to use,for example, water or physiologically acceptable organic solvents suchas alcohols (e.g., ethanol, propanol, isopropanol, 1,2-propylene glycol,polyglycols and their derivatives, fatty alcohols, partial esters ofglycerol), oils (e.g., peanut oil, olive oil, sesame oil, almond oil,sunflower oil, soya bean oil, or castor oil), paraffins, dimethylsulphoxide, triglycerides and the like. In the case of liquidformulations, the following substances may be used as stabilizers orsolubilizers: lower aliphatic mono- and multivalent alcohols with 2-4carbon atoms such as ethanol, n-propanol, glycerol, polyethylene glycolswith molecular weights between 200-600 (e.g., 1 to 40% aqueoussolution), gum acacia, guar gum, or other suspension agents selectedfrom the hydrocolloids.

In addition to liquid formulations for oral delivery, solutions orsuspensions used for non-oral delivery (e.g., parenteral, intradermal,subcutaneous, or topical) can include any of the following components: asterile diluent such as water, saline solution, fixed oil, polyethyleneglycol, glycerine, propylene glycol, dimethyl acetamide or othersynthetic solvents; antimicrobial agents such as benzyl alcohol andmethyl parabens; antioxidants such as ascorbic acid and sodiumbisulfite; chelating agents such as ethylenediaminetetraacetic acid(EDTA); buffers such as acetates, citrates and phosphates; and agentsfor the adjustment of tonicity such as sodium chloride or dextrose.Parenteral preparations can be enclosed in ampules, disposable syringesor single or multiple dose vials made of glass, plastic or othersuitable material.

In instances in which the free beta-alanine or a biological sourcethereof exhibit insufficient solubility, methods for solubilizing freebeta-alanine or a biological source thereof may be used. Such methodsare known to those of skill in the art and include, but are not limitedto, using co-solvents such as dimethylsulfoxide (DMSO),dimethylacetamide, using surfactants such as TWEEN®, or by dissolvingthe beta-alanine in aqueous sodium bicarbonate.

In one embodiment, beta-alanine or a biological source thereof can beformulated in a liposomal suspension (e.g., tissue-targeted liposomes)for delivery to a human or an animal. A liposomal suspension can beprepared according to methods known to those skilled in the art. See,for example, U.S. Pat. No. 4,522,811. Briefly, liposomes such asmultilamellar vesicles (MLV's) may be formed by drying down eggphosphatidyl choline and brain phosphatidyl serine (e.g., at a 7:3 molarratio) on the inside of a flask. A solution of beta-alanine or abiological source thereof in phosphate buffered saline lacking divalentcations (PBS) is added and the flask shaken until the lipid film isdispersed. The resulting vesicles are washed to remove un-encapsulatedbeta-alanine, pelleted by centrifugation, and then resuspended in PBS.

Pharmaceutically acceptable carriers are known in the art, and theirselection usually is dependent upon the intended route of delivery.Examples of carriers useful in solid and aqueous dispersions accordingto the compositions and methods provided herein include, but are notlimited to, water-soluble polymers such as guar gum, glucommannan,psyllium, gum acacia, polyethylene glycol, polyvinylpyrrolidone,hydroxypropyl methylcellulose, and other cellulose ethers such asmethylcellulose, and sodium carboxymethylcellulose. Additional carriersfor a liquid composition include, for example, water, saline, aqueousdextrose, glycerol, glycols, ethanol, and the like.

Along with the beta-alanine or a biological source thereof, thecomposition can contain any number of additional components. Forexample, a composition containing beta-alanine also can include: adiluent such as lactose, sucrose, dicalcium phosphate, or carboxymethylcellulose; a lubricant, such as magnesium stearate, calciumstearate and talc; and/or a binder such as starch, natural gums, such asgum acaciagelatin, glucose, molasses, polvinylpyrrolidine, cellulosesand derivatives thereof, povidone, crospovidones and other such bindersknown to those of skill in the art. Compositions of the inventionfurther can include disintegrating agents, coloring agents (e.g., any ofthe approved certified water soluble FD&C dyes), flavoring agents, andwetting agents. Flavoring and sweetening agents are useful, as arepreservatives, stabilizers, buffer substances and antioxidants.

The pharmaceutical composition disclosed herein also may contain furthersubstances such as emulsifying agents, solubilizing agents, pH bufferingagents, and the like (e.g., acetate, sodium citrate, cyclodextrinederivatives, sorbitan monolaurate, triethanolamine sodium acetate,triethanolamine oleate, and other such agents). Solvents includeglycerin, sorbitol, ethyl alcohol and syrup; examples of preservativesinclude glycerin, methyl and propylparaben, benzoic add, sodium benzoateand alcohol; examples of non aqueous liquids utilized in emulsionsinclude mineral oil and cottonseed oil; examples of emulsifying agentsinclude gelatin, acacia, tragacanth, bentonite, and surfactants such aspolyoxyethylene sorbitan monooleate; suspending agents include sodiumcarboxymethylcellulose, pectin, tragacanth, Veegum and acacia;sweetening agents include sucrose, syrups, glycerin and artificialsweetening agents such as saccharin; and wetting agents includepropylene glycol monostearate, sorbitan monooleate, diethylene glycolmonolaurate and polyoxyethylene lauryl ether.

Liquid pharmaceutically-administrable compositions can, for example, beprepared by dissolving, dispersing, or otherwise mixing freebeta-alanine or a biological source thereof and optional pharmaceuticaladjuvants in a carrier, such as, for example, water, saline, aqueousdextrose, glycerol, glycols, ethanol, and the like, to thereby form asolution or suspension. In addition, tablets and capsules formulationsmay be coated as known by those of skill in the art in order to modifyor sustain dissolution of the active ingredient. Thus, for example, theymay be coated with a conventional coating such as phenylsalicylate,waxes and cellulose acetate phthalate, or with another enteric coatingthat maintains its integrity in the stomach and provides sustainedrelease of the beta-alanine. Methods of preparing pharmaceuticalcompositions are known or will be apparent to those skilled in this art.See, for example, Remington's Pharmaceutical Sciences, Mack PublishingCompany, Easton, Pa., 15th Edition, 1975.

C. Sustained Release Formulations

The preferred formulation for the compositions containing freebeta-alanine or a biological source thereof as described herein is asustained release formulation. For the purposes of the presentinvention, the following terms may be considered to be substantiallyequivalent to ‘sustained release’: continuous release, controlledrelease, delayed release, depot, gradual release, long-term release,programmed release, prolonged release, proportionate release, protractedrelease, repository, retard, slow release, spaced release, time coat,timed release, delayed action, extended action, layered-time action,long-acting, prolonged action, repeated action, slowing acting,sustained action, sustained-action medications, and extended release.Further discussions of these terms may be found in Lesczek Krowczynski,Extended-Release Dosage Forms, 1987 (CRC Press, Inc.).

The concentrations of free beta-alanine or a biological source thereofin the compositions disclosed herein are effective for increasing theanaerobic working capacity of a tissue by providing an amount of freebeta-alanine to blood or blood plasma effective to increase thesynthesis of beta-alanylhistidine peptides, particularly carnosine, inmuscle and other tissues. A sustained release formulation ofbeta-alanine is administered in an amount effective to attenuate themetabolic acidosis encountered during high-intensity exercise and thusdelay the onset of fatigue during anaerobic exercise without inducingsymptoms of paraesthesia.

The concentration of free beta-alanine or a biological source thereof inthe pharmaceutical composition will depend on the particularformulation, the rate of absorption, the inactivation and excretionrates of the beta-alanine, the physicochemical characteristics of thebeta-alanine, and the dosage schedule as well as other factors known tothose of skill in the art. The contemplated compositions, however, maycontain 0.001% up to 100% of free beta-alanine or a biological sourcethereof (e.g., 0.1-85% or 75-95%).

The free beta-alanine or a biological source thereof is included in thepharmaceutically acceptable carrier in an amount sufficient to exert atherapeutically useful effect in the absence of undesirable sideeffects. The therapeutically effective concentration may be determinedempirically by testing an amount of beta-alanine in in vitro and in vivosystems such as those described herein and then extrapolating therefromfor dosages in humans.

Typically, a therapeutically effective dosage should produce a blood,plasma or serum concentration of beta-alanine or an equivalent amount ofa biological source thereof to muscle or other tissue of from about 0.1μg/ml to about 50-100 μg/ml. The pharmaceutical compositions typicallyshould provide a dosage of from about 0.001 mg to about 2000 mg ofbeta-alanine per kg of body weight per day. Pharmaceutical dosage unitforms are prepared to provide from about 1 mg to about 10,000 mg (e.g.,from about 10 to about 5,000 mg) of the beta-alanine per dosage unitform.

The free beta-alanine dosage can be between about 1 milligram (mg) andabout 200 mg per kilogram (kg) body weight, or the dose of a biologicalsource thereof (e.g., a peptide of beta-alanine (e.g. carnosine)) can bebetween about 2.5 mg and about 500 mg per kg body weight. By way ofexample, suitable controlled release dosages in an 80 kg person per daycan be between 0.08 grams to 16.0 grams of free beta-alanine or anequivalent amount of a biological source of beta-alanine. In one aspect,the total amount of free beta-alanine administered in a controlledrelease dosage per day can be at least 200 mg, from 200 mg to 5 g, from5 g to 10 g, or from 5 g to 16 g or more per day for a human. A singledose of active ingredient, e.g., free beta-alanine or a biologicalsource thereof, may be formulated to be in the amount about 200, 400,800, 1600, 3200, 6400 mg or more.

For humans and animals, the compositions disclosed herein can be, forexample, (a) 1% to 99% by weight of free beta-alanine or a biologicalsource thereof and 0% to 98% by weight of water; (b) 1% to 98% by weightof free beta-alanine or a biological source thereof and 0% to 97% byweight of water; (c) 1% to 20% by weight of free beta-alanine or abiological source thereof, 9% to 99% by weight of glucose or othersimple carbohydrate and 0% to 60% by weight of water; or (d) 1% to 20%by weight of free beta-alanine or 1% to 20% by weight of a peptide ofbeta-alanine, 9% to 99% by weight of glucose or other simplecarbohydrate and 0% to 60% by weight of water.

A particular dosage of beta-alanine can be administered in a single doseor can be divided into a number of smaller doses to be administered atparticular intervals of time. It is to be noted that concentrations anddosage values may also vary with the severity of the condition to bealleviated. It is to be further understood that for any particularsubject, specific dosage regimens should be adjusted over time accordingto the individual need and the professional judgment of the personadministering or supervising the administration of the compositions, andthat the concentration ranges set forth herein are exemplary only andare not intended to limit the scope or practice of the claimedcompositions.

The pharmaceutically therapeutically active free beta-alanine,biological sources of beta-alanine, and derivatives, analogues orconjugates thereof are typically formulated and administered in unitdosage forms or multiple dosage forms. Unit dose forms as used hereinrefers to physically discrete units suitable for human and animalsubjects and packaged individually as is known in the art. Each unitdose contains a predetermined quantity of therapeutically activebeta-alanine sufficient to produce the desired therapeutic effect inassociation with the required pharmaceutical carrier, vehicle ordiluent. Unit dose forms may be administered in fractions or multiplesthereof. A multiple dose form generally is a plurality of identical unitdosage forms packaged in a single container to be administered insegregated unit dose form. Examples of unit dose forms include ampulesand syringes and individually packaged tablets or capsules, and examplesof multiple dose forms include vials, bottles of tablets or capsules,and bottles of pints or gallons.

As used herein, sustained release can mean any one of a number ofextended release dosage forms. The various sustained releasetechnologies cover a very broad spectrum of drug dosage forms. Sustainedrelease technologies include, but are not limited to, physical systemsas well as chemical systems.

Physical systems include, but are not limited to, reservoir systems withrate-controlling membranes such as encapsulation (e.g., micro- andmacro-) and membrane systems; reservoir systems without rate-controllingmembranes such as hollow fibers, ultra macroporous cellulose triacetate,and porous polymeric substrates and foams; monolithic systems includingthose systems physically dissolved in non-porous, polymeric, orelastomeric matrices (e.g., non-erodible, erodible, environmental agentingression, and degradable) and materials physically dispersed innon-porous, polymeric, or elastomeric matrices (e.g., non-erodible,erodible, environmental agent ingression, and degradable); laminatedstructures including reservoir layers chemically similar or dissimilarto outer control layers; and other physical methods such as osmoticpumps or adsorption onto ion-exchange resins.

Chemical systems include, but are not limited to, chemical erosion ofpolymer matrices (e.g., heterogeneous or homogeneous erosion) orbiological erosion of a polymer matrix (e.g., heterogeneous, orhomogeneous). Hydrogels may also be employed as described in “ControlledRelease Systems: Fabrication Technology”, Vol. II, Chapter 3; pages41-60; “Gels For Drug Delivery”, Edited By Hsieh, D.

There are a number of sustained release drug formulations that have beendeveloped. These include, but are not limited to, microencapsulatedpowders; osmotic pressure-controlled gastrointestinal delivery systems;hydrodynamic pressure-controlled gastrointestinal delivery systems;membrane permeation-controlled gastrointestinal delivery systems, whichinclude microporous membrane permeation-controlled gastrointestinaldelivery devices; gel diffusion controlled gastrointestinal deliverysystems; and ion-exchange-controlled gastrointestinal delivery systems,which include cationic and anionic drugs. In one embodiment, thesustained release system can be an oil-microencapsulated sustainedrelease powder dosage form that can be mixed with liquid and consumed asa drink mix beverage. See, also, sustained release formulationsdeveloped by Rohm Hass (Philadelphia, Pa.), as well as representativeU.S. patents that disclose sustained release formulations (e.g., U.S.Pat. Nos. 6,696,500; 6,756,049; 6,919,372; 6,992,065; and 7,048,947).

Furthermore, many of the conventional pharmaceutical binders, excipientsand additives, when used in sufficient quantities, can act ascontrolled-release polymers and produce a sustained release effect.These include, but are not limited to, gelatin, natural sugars such asraw sugar or lactose, lecithin, mucilage, plant gums, pectins or pectinderivatives, algal polysaccharides, glucomannan, agar and lignin, guargum, locust bean gum, acacia gum, xanthan gum, carrageenan gum, karayagum, tragacanth gum, ghatti gum, starches (e.g., corn starch oramylose), dextran, polyvinyl pyrrolidone, polyvinyl acetate, gum arabic,alginic acid, tylose, talcum, lycopodium, silica gel (e.g., colloidal),cellulose and cellulose derivatives (e.g., cellulose ethers, celluloseethers in which the cellulose hydroxy groups are partially etherifiedwith lower saturated aliphatic alcohols and/or lower saturated,aliphatic oxyalcohols; e.g., methyloxypropyl cellulose, methylcellulose, hydroxypropyl methyl cellulose, hydroxypropyl methylcellulose phthalate, cross-linked sodium carboxymethylcellulose,cross-linked hydroxypropylcellulose, high-molecular weighthydroxymethylpropylcellulose, carboxymethyl-cellulose, low-molecularweight hydroxypropylmethylcellulose medium-viscosityhydroxypropylmethylcellulose hydroxyethylcellulose,hydroxypropylcellulose, hydroxypropylmethylcellulose, sodiumcarboxymethylcellulose, alkylcelluloses, ethyl cellulose, celluloseacetate, cellulose propionate (low, medium or high molecular weight),cellulose acetatepropionate, cellulose acetate butyrate, cellulosetriacetate, methyl cellulose, hydroxypropyl cellulose, orhydroxypropylmethyl cellulose), fatty acids as well as magnesium,calcium or aluminum salts of fatty acids with 12 to 22 carbon atoms, inparticular saturated fatty acids (e.g., stearates such as magnesiumstearate), polycarboxylic acids, emulsifiers, oils and fats, inparticular vegetable (e.g., peanut oil, castor oil, olive oil, sesameoil, cottonseed oil, corn oil, wheat germ oil, sunflower seed oil, codliver oil, in each case also optionally hydrated); glycerol esters andpolyglycerol esters of saturated fatty acids (e.g., C₁₂H₂₄O₂ toC₁₈H₃₆O₂) and their mixtures, it being possible for the glycerol hydroxygroups to be totally or also only partly esterified (e.g., mono-, di-and triglycerides); high melting point hydrogenated vegetable oilssuitable for microencapsulation; pharmaceutically acceptable mono- ormultivalent alcohols and polyglycols such as polyethylene glycol andderivatives thereof, esters of aliphatic saturated or unsaturated fattyacids (e.g., 2 to 22 carbon atoms, e.g., 10-18 carbon atoms) withmonovalent aliphatic alcohols (e.g., 1 to 20 carbon atoms) ormultivalent alcohols such as glycols, glycerol, diethylene glycol,pentacrythritol, sorbitol, mannitol and the like, which may optionallyalso be etherified, esters of citric acid with primary alcohols, aceticacid, urea, benzyl benzoate, dioxolanes, glyceroformals,tetrahydrofurfuryl alcohol, polyglycol ethers with C₁-C₁₂ alcohols,dimethylacetamide, lactamides, lactates, ethylcarbonates, silicones(e.g., medium-viscous polydimethyl siloxanes), calcium carbonate, sodiumcarbonate, calcium phosphate, sodium phosphate, magnesium carbonate andthe like.

In addition, coating agents may be used to create or enhance thesustained release features of a pharmaceutical composition. For example,plasticizers that are useful as coating agents include, but are notlimited to, citric and tartaric acid esters (acetyl-triethyl citrate,acetyl tributyl-, tributyl-, triethyl-citrate); glycerol and glycerolesters (glyceroldiacetate, -triacetate, acetylated monoglycerides,castor oil); phthalic acid esters (dibutyl-, diamyl-, diethyl-,dimethyl-, dipropyl-phthalate), di-(2-methoxy- or2-ethoxyethyl)-phthalate, ethylphthalyl glycolate, butylphthalylethylglycolate and butylglycolate; alcohols (propylene glycol, polyethyleneglycol of various chain lengths); adipates (diethyladipate,di(2-methoxy- or 2-ethoxyethyl)-adipate; benzophenone; diethyl- anddiburylsebacate; dibutylsuccinate; dibutyltartrate; diethylene glycoldipropionate; ethyleneglycol diacetate, -dibutyrate, -dipropionate;tributyl phosphate, tributyrin; polyethylene glycol sorbitan monooleate;polysorbates such as Polysorbar 50; and sorbitan monooleate.

Methods of making sustained release pharmaceutical compositions areknown in the art. For example, coating agents such as ethylcellulose andhydroxypropyl methylcellulose can be mixed together and sprayed ontobeta-alanine or a biological source thereof in a fluid bed granulator.Another method of making a sustained release pharmaceutical compositionemploys mixtures of a high temperature melted vegetable oil (e.g.,having an iodine value maximum of about 5 and a melting point of about145° F.) with a cellulose ether such as ethylcellulose. This combinationcan be processed in a vertical or horizontal high intensity mixer or ablender that is jacketed so as to allow a hot water bath to circulatearound the mixer to elevate the temperature of the oil to the meltingpoint. Powdered beta-alanine or a biological source thereof is thenmixed with the molten oil until complete coverage is achieved (about5-10 minutes), cooled, and the ethylcellulose sprayed onto theparticles. The finished product is a microencapsulated, free-flowingsustained-release powder with an extended release profile. See, also,U.S. Pat. Nos. 6,835,397; 6,013,286; and 5,190,775 for methodsdescribing controlled release of encapsulated bioactive substances.

In the compositions, effective concentrations of free beta-alanine or abiological source thereof are mixed with a suitable pharmaceuticalcarrier or vehicle. The free beta-alanine or a biological source thereofmay be derivatized as the corresponding salts, esters, enol ethers oresters, acids, bases, solvates, hydrates or pro-drugs prior toformulation, as is known in the art.

D. Dietary Supplements

Dietary supplements formulated for the sustained release of freebeta-alanine or a biological source thereof are provided. The dietarysupplements may comprise a food product or a liquid product.

1. Solid Food Products

A dietary supplement formulated as a food product can be in a solid oran edible suspension. Solid food products may comprise chewable oredible bars, cookies, biscuits, lozenges, chewing gum, or ediblesuspension. In one embodiment, the dietary supplement containingfree-beta alanine or a biological source thereof is a solid food productin the form of a high-energy multi-saccharide edible bar containing thesubstance galactose.

In one embodiment, an edible food bar includes a saccharide componentincluding 3 to 37% weight/weight (w/w) galactose, 0.1 to 75% w/w freebeta-alanine or a biological source thereof, and optional furtheringredients selected from amino acids, carbohydrates, fiber and fat, andother ingredients such as creatine and beta-alanylhistidine peptides(e.g. carnosine, anserine, and/or balenine). The amount of galactose canbe, for example, 5 to 20% w/w (e.g., 5 to 15% w/w). The saccharidecomponent also can include glucose. In one embodiment, equal amounts ofglucose and galactose are provided. In, another embodiment, the amountof galactose is greater than the amount of glucose.

The invention generally includes the inclusion of free beta-alanine or abiological source thereof with other ingredients to give a bar, drink orother type of food that can be low in glucose and can have a lowglycemic index. The use of galactose affords several advantages. Forexample, galactose is not insulogenic; that is, galactose does notitself induce an insulin response. Thus, its use is associated with lessof an insulin response than the equivalent mass of glucose. Products inaccordance with this invention may be used by diabetics or persons whoare intolerant to lactose. Galactose can be used rapidly by the liverfor synthesis of glycogen or glucose, and it is less likely to causedental erosion than other sugars.

Some embodiments of solid food product compositions containing freebeta-alanine incorporate 0.1 to 50% free beta-alanine or a biologicalsource thereof. In order to prevent unwanted adverse side effects, it isdesired that the free beta-alanine or a biological source thereofexhibits sustained or delayed release from the food product. Forexample, the free beta-alanine or a biological source thereof may becontained within a food product and separated from any moist ingredientsby, for example, formulation in layers. Alternatively, the freebeta-alanine or a biological source thereof may be embodied within thematrix of the food itself, wherein the nature of the matrix delaysdissolution of the food within the stomach. Additionally, beta-alaninecan be encapsulated as granules or powder in a dry water-imperviousshell (e.g., micro-encapsulation) and used in a food product. In thisway, the free beta-alanine can be slowly released in the stomach bydissolution.

Use of fiber in a food product (e.g., an energy bar) is advantageousbecause different fiber products influence the release of sugars, affectthe binding of various components and advantageously delay digestion. Anamount of between 0 to 5% (or more) w/w sugars may be used in a foodproduct containing beta-alanine or a biological source thereof.Furthermore, a composition containing beta-alanine or a biologicalsource thereof may be absorbed onto the fiber prior to the manufactureof the food product. In this way, the absorption of the beta-alanineinto the body may be retarded.

In a layered arrangement, the free beta-alanine powder or aggregate maybe covered with a hard dry sugar layer. Alternatively or in addition, achocolate containing layer may be used as a moisture barrier. A freebeta-alanine containing layer may be provided as a laminar cylindricallayer disposed within the bar as the inner layer. Alternatively or inaddition, ingredients such as fiber, nuts and dried fruit may be layeredonto the layer-containing free beta-alanine to form a laminate. Such alayer may incorporate syrup so as to form a composite-type layer.

In alternative food products, beta-alanine powder or aggregate may becovered with a hard sugar mixture, chocolate or both to form particleswith a mass of 100 to 500 mg. These can be combined with the remainingingredients to form dispersed units within the matrix, for example, aschocolate chips within a cookie. Such an encapsulated arrangement mayincorporate layers as previously described.

2. Edible Suspensions

In another embodiment, dietary supplements containing free-beta alanineor a biological source of beta-alanine are food products wherein thefree beta-alanine or a biological source of beta-alanine is suspended inan edible supporting matrix to form an edible suspension. The term“suspension” is intended to mean compositions containing freebeta-alanine or a biological source of beta-alanine as provided herein,comprise flee beta-alanine in solid form (e.g. as crystals, power or thelike), distributed within an edible viscous liquid or semi-liquid, or asolid, supporting matrix, typically such that settling (under theinfluence of gravity) of the solid beta-alanine is inhibited orprevented.

The composition may be provided in solid, liquid or semi-liquid form(e.g. as a drink, soup or yogurt). Preferably, the free beta-alanine isdistributed substantially evenly throughout the supporting matrix (byhomogenizing in some manner e.g. by stirring, blending or the like),which may be accomplished manually (e.g. by the consumer) and/ormechanically at the time the composition is prepared.

Conveniently, the food product is an otherwise conventional food productsupplemented with free beta-alanine or a biological source ofbeta-alanine such that beta-alanine becomes suspended in the foodstuff.Examples of foodstuffs that may represent suitable supporting matricesfor the composition of the invention include spreadable solids such asdairy or cheese spreads, margarines, meat and fish pastes and spreadsand the like. Other convenient supporting matrices are those containingsugars or other carbohydrates, such as liquid (“runny”) or solid (“set”)honey, molasses, syrup (e.g. corn syrup, glucose syrup), treacle or gelsof any description, foods made viscous by cooling including ice-cream,and foods made viscous by cooking and baking, e.g. muffins, pies, tarts,cakes, biscuits and cereal flakes.

If desired, the viscosity of the edible matrix and/or the composition asa whole, may be increased by the addition of viscosifiers, gellingagents and the like. Such components are well-known in the food industryand include, for example, plant-derived polysaccharides, gums and thelike such as galactomannans, dextrans, guar gum, locust bean gum and soon. Such viscosifiers, gels and the like may form the supporting matrix,if desired. One representative edible matrix comprises a gel preparedfrom concentrated Aloe Vera extract: a smooth creamy paste, which, forexample, can be packaged in a squeezable tube.

The composition may comprise one or more further components to improveits palatability, stability, flavor or nutritive quality. These furthercomponents may include electrolytes, vitamins (e.g., vitamin E, vitaminC, thiamin, riboflavin, niacin, vitamin B6, folic acid, vitamin B12,biotin, and pantothenic acid), lipids, carbohydrates (e.g., starchand/or sugars, e.g., glucose, fructose, sucrose, and maltrose), aminoacids, trace elements, colorings, flavors, artificial sweeteners,natural health improving substances, anti-oxidants, stabilizers,preservatives, and buffers. The composition may be unflavored or havethe normal flavor of the matrix. Alternatively, one or more flavors maybe added (e.g. fruit, cheese or fish flavor).

Other ingredients that can be included in the presently disclosedsustained release beta-alanine compositions can include, for example,anti-oxidants, alpha-lipoic acid, tocotrienols, N-acetylcysteine,co-enzyme Q-10, extracts of rosemary such as carnosol, botanicalanti-oxidants such as green tea polyphenols, grape seed extract, COX-1type inhibitors such as resveratrol, ginkgo biloba, and garlic extracts.Other amino acids such as L-cysteine or L-citrulline may be added.Combination with an acetylcholine precursor such as choline chloride orphosphatidylcholine may be desirable, for example, to enhancevasodilation. It is to be understood that such combination therapyconstitutes a further aspect of the compositions and methods oftreatment provided herein.

Artificial sweeteners which can be used include Aspartame, Acesulfam K,Saccharin and Cyclamate. Almost any desired flavoring can be added suchas fruity flavors such as berry, lemon, orange, papaya and grapefruit.Citric acid may be used as an acidulant and citrate (e.g. sodiumcitrate) as a buffering agent. Also, other natural health improvingsubstances such as Pan D'Arco tea, Ginseng, Suma tea, Ginkgo, bee pollenand myrrh may be added in physiologically active amounts. Preservativessuch as potassium benzoate and/or potassium sorbate can be included.Coloring can be included such as cold water soluble colorant such asbeta-carotene. Other suitable colorings, however, will be apparent tothose skilled in the art. A clouding agent may be included in thecomposition, if desired, to improve the appearance of the composition.

Mineral and trace elements also can be added in any type or form whichis suitable for human consumption. It is convenient to provide thecalcium and potassium in the form of their gluconates, phosphates orhydrogen phosphates, and magnesium as the oxide or carbonate, chromiumas chromium picolinate, selenium as sodium selenite or selenate, andzinc as zinc gluconate. Representative amounts include sodium at 400mg/liter, calcium at 100 mg/liter, chloride at 600 mg/liter, potassiumat 200 mg/liter, magnesium at 75 mg/liter and phosphorus at 50 mg/liter,chromium at 125 μg/liter, selenium at 125 μg/liter and zinc at 15mg/liter.

In accordance with the present invention, there may be employedconventional molecular biology, microbiology, biochemical, andrecombinant DNA techniques within the skill of the art. Such techniquesare explained fully in the literature. The invention will be furtherdescribed in the following examples, which do not limit the scope of theinvention described in the claims.

EXAMPLES Example 1 Edible Gel Formulation for the Sustained Release ofFree Beta-Alanine

To prepare an edible gel containing free beta-alanine the contents oftwo packets of commercially available strawberry flavored gelatin gelwere dissolved in 200 ml of boiling water. The contents were stirredcontinuously until the gel was completely dissolved. The gel was furtherfortified with the addition of a sachet of gelatin capsules. To this wasadded 80 g free beta-alanine (in a final volume of approximately 300ml). The concentrated gel solution was poured into a 12×12 cm flat mouldto a depth of approximately 2 cm and allowed to set to a stiff gel at 4°C. One square piece of gel (approximately 2×2×2 cm in size) containedapproximately 2 g of beta-alanine.

When the piece of gel containing 2 g beta-alanine was ingested (eitheras two half pieces), symptoms of paraesthesia were mild in the following20 minutes, and in some subjects could not be detected at all. Incontrast, symptoms of paraesthesia were detected in subjects thatingested 2 g free beta-alanine dissolved in water. Thus the sustainedrelease of free beta-alanine mitigates the symptoms of paraesthesia thatare felt when equivalent amounts are administered in a single bolusdose.

Example 2 Dissolution of Beta-Alanine Capsules

Samples. Beta-alanine capsules produced by Balchem Encapsulates (NewHampton, N.Y.; see, for example, U.S. Pat. No. 6,835,397) were filledwith lipid coated granules of beta-alanine. Samples were identified asRIA-1743 60%, 70%, 80%, 90% and RIA-1744 60%, 70%, 80%, 90% based on theamount of beta-alanine present.

Experimental Testing. Testing was performed by Natural AlternativeInc.'s Research & Development Laboratory utilizing the Logan InstrumentsDISSO III, USP test apparatus 3. Dissolution was carried out in 0.1 Nhydrochloric acid for the length of six hours. Samples were taken at 1,2, 3, and 6 hours.

Screens used for the first analysis were single size 40 mesh for the topand bottom caps. The second analysis with RIA-1744 applied two meshscreens for the top and bottom caps: a size 40 mesh screen and a secondmesh screen (with slightly larger hole size) were combined. Mesh screenswere combined as an effort to contain the granules inside thereciprocating cylinders. Granules were released from the capsulesfollowing capsule shell disintegration. Quantitation was performed usingHPLC analysis with evaporative light scattering detection.

Results. Tables 1 and 2 show the results using the single mesh screenssize 40, and Table 3 shows the analysis of RIA-1744 with double meshscreens. The data show that the 80% and 90% samples of bothformulations, RIA-1743 and RIA-1744, demonstrate little sustainedrelease properties. The RIA-1744 60% and 70% formulation demonstratedsustained release properties in both analyses. The 60% formulation hadeven less beta-alanine dissolved at the end of six hours.

TABLE 1 RIA-1743 % Dissolved Hour 1 2 3 6 60% 0.0 8.3 8.8 10.0 70% 11.312.4 14.1 17.2 80% 27.9 35.2 44.6 44.8 90% 88.9 93.4 76.8 95.9

TABLE 2 RIA-1744 % Dissolved Hour 1 2 3 6 60% 18.6 33.9 49.3 74.9 70%26.3 46.8 62.6 81.2 80% 88.1 90.3 73.2 91.3 90% 96.1 94.6 93.7 93.5

TABLE 3 RIA-1744 % Dissolved Hour 1 2 3 6 60% 20.8 33.1 46.4 80.4 70%26.2 46.7 58.0 84.8 80% 86.4 94.4 92.5 98.1

Example 3 Sustained Release Formulation Compared to a Non-SustainedRelease Formulation

Two subjects were given 1.75 g beta-alanine dissolved in water(formulated as a drink) or 5 capsules, each containing 0.35 gbeta-alanine in a delayed release formulation. Subjects reportedsymptoms of paraesthesia affecting the face, scalp, neck, arms, handsand buttocks with 1.75 g of beta-alanine administered in a drink, butnot with 1.75 g administered in the delayed release capsule formulation.Results (mean±SE) are shown in FIG. 1.

Example 4 Sustained Release Formulations

Subjects were given capsules containing 400 mg beta-alanine but asformulations composed of 100%, 90%, 80%, 70% and 60% beta-alanine, withthe balance comprising a lipid mixture to delay dissolution. Twodifferent lipid mixtures were used, referred to herein as mixtures “A”and “B”.

FIG. 2 shows the compositions of “A” and “B” that contained the highestbeta-alanine concentration in which paraesthesia symptoms were notrecorded. Each column of figures denotes (from top to bottom): subjectnumber; gender; weight (kg); and number of 400 mg capsules ingested.

Subjects 1-6 did not record paraesthesia symptoms even when the capsuleswere entirely (i.e., 100%) beta-alanine. Subjects 1-4 took 5 capsules ateach dose and subjects 5-6 took 4 capsules at each dose. Subjects 7-10went to a lower % of beta-alanine before paraesthesia symptomsdisappeared completely, more so when taking mixture “B” compared tomixture “A”. Each of subjects 7-10 took 4 capsules at each dose.Subjects 11-14 were more sensitive to the paraesthesia effects ofbeta-alanine that the other subjects, recording the highest gradesymptoms at 100% beta-alanine. No paraesthesia symptoms were experiencedby subject 11 with 60% beta-alanine in either mixture “A” or “B”.Subjects 12-13 still experienced mild paraesthesia symptoms at 60%beta-alanine in either mixture and were, therefore, projected as beingsymptom free at 50% in either mixture. Subjects 13 and 14 took just 3capsules at each dose.

Other Embodiments

It is to be understood that while the invention has been described inconjunction with the detailed description thereof, the foregoingdescription is intended to illustrate and not limit the scope of theinvention, which is defined by the scope of the appended claims. Otheraspects, advantages, and modifications are within the scope of thefollowing claims.

The invention claimed is:
 1. A dietary supplement comprising an amountof a microencapsulated, sustained release free beta-alanine, or a saltthereof in a microencapsulated, sustained release formulation, whereinfollowing ingestion of the dietary supplement by a human a concentrationof beta-alanine in the human's blood does not exceed 200 μmol/L within15 minutes following ingestion, wherein the free beta-alanine or saltthereof, in the sustained release formulation of the dietary supplementis provide in an amount ranging from about 0.1 to about 10.0 grams. 2.The dietary supplement of claim 1 wherein the free beta-alanine, or saltthereof, in the sustained release formulation is provided in an amountranging from about 1.0 to about 10.0 grams.
 3. The dietary supplement ofclaim 1 wherein the free beta-alanine, or salt thereof, in the sustainedrelease formulation is provided in an amount ranging from about 1.0 toabout 9.0 grams.
 4. The dietary supplement of claim 1 wherein the freebeta-alanine, or salt thereof, in the sustained release formulation isprovided in an amount ranging from about 2.0 to about 8.0 grams.
 5. Thedietary supplement of claim 1 wherein the free beta-alanine, or saltthereof, in the sustained release formulation is provided in an amountranging from about 3.0 to about 7.0 grams.
 6. The dietary supplement ofclaim 1 wherein the free beta-alanine, or salt thereof, in the sustainedrelease formulation is provided in an amount ranging from about 4.0 toabout 6.0 grams.
 7. The dietary supplement of claim 1 wherein thesustained release formulation is in a solid, liquid or semi-liquid.
 8. Adietary supplement comprising an amount of microencapsulated, sustainedrelease free beta-alanine, or a salt thereof in a microencapsulated,sustained release formulation, wherein following ingestion of thedietary supplement by a human a concentration of beta-alanine in thehuman's blood does not exceed 375 μmol/L within 30 minutes followingingestion, wherein the free beta-alanine or salt thereof, in thesustained release formulation of the dietary supplement is provided inan amount ranging from about 0.1 to about 10.0 grams.
 9. The dietarysupplement of claim 8 wherein the free beta-alanine, or salt thereof, inthe sustained release formulation is provided in an amount ranging fromabout 1.0 to about 10.0 grams.
 10. The dietary supplement of claim 8wherein the free beta-alanine, or salt thereof, in the sustained releaseformulation is provided in an amount ranging from about 1.0 to about 9.0grams.
 11. The dietary supplement of claim 8 wherein the freebeta-alanine, or salt thereof, in the sustained release formulation isprovided in an amount ranging from about 2.0 to about 8.0 grams.
 12. Thedietary supplement of claim 8 wherein the free beta-alanine, or saltthereof, in the sustained release formulation is provided in an amountranging from about 3.0 to about 7.0 grams.
 13. The dietary supplement ofclaim 8 wherein the free beta-alanine, or salt thereof, in the sustainedrelease formulation is provided in an amount ranging from about 4.0 toabout 6.0 grams.
 14. The dietary supplement of claim 8 wherein thesustained release formulation is in a solid, liquid or semi-liquid. 15.A dietary supplement comprising an amount of a microencapsulated,sustained release free beta-alanine, or a salt thereof in amicroencapsulated, sustained release formulation, wherein followingingestion of the dietary supplement by a human a concentration ofbeta-alanine in the human's blood does not exceed 300 μmol/L within 45minutes following ingestion, wherein the free beta-alanine or saltthereof, in the sustained released formulation of the dietary supplementis provided in an amount ranging from about 0.1 to about 10.0 grams. 16.The dietary supplement of claim 15 wherein the free beta-alanine, orsalt thereof, in the sustained release formulation is provided in anamount ranging from about 1.0 to about 10.0 grams.
 17. The dietarysupplement of claim 15 wherein the free beta-alanine, or salt thereof,in the sustained release formulation is provided in an amount rangingfrom about 1.0 to about 9.0 grams.
 18. The dietary supplement of claim15 wherein the free beta-alanine, or salt thereof, in the sustainedrelease formulation is provided in an amount ranging from about 2.0 toabout 8.0 grams.
 19. The dietary supplement of claim 15 wherein the freebeta-alanine, or salt thereof, in the sustained release formulation isprovided in an amount ranging from about 3.0 to about 7.0 grams.
 20. Thedietary supplement of claim 15 wherein the free beta-alanine, or saltthereof, in the sustained release formulation is provided in an amountranging from about 4.0 to about 6.0 grams.
 21. The dietary supplement ofclaim 15 wherein the sustained release formulation is in a solid, liquidor semi-liquid.
 22. A dietary supplement comprising an amount of amicroencapsulated, sustained release free beta-alanine, or a saltthereof in a microencapsulated, sustained release formulation, whereinfollowing ingestion of the dietary supplement by a human a concentrationof beta-alanine in the human's blood does not exceed 100 μmol/L within75 minutes following ingestion, wherein the free beta-alanine or saltthereof, in the sustained release formulation of the dietary supplementis provided in an amount ranging from about 0.1 to about 10.0 grams. 23.The dietary supplement of claim 22 wherein the free beta-alanine, orsalt thereof, in the sustained release formulation is provided in anamount ranging from about 1.0 to about 10.0 grams.
 24. The dietarysupplement of claim 22 wherein the free beta-alanine, or salt thereof,in the sustained release formulation is provided in an amount rangingfrom about 1.0 to about 9.0 grams.
 25. The dietary supplement of claim22 wherein the free beta-alanine, or salt thereof, in the sustainedrelease formulation is provided in an amount ranging from about 2.0 toabout 8.0 grams.
 26. The dietary supplement of claim 22 wherein the freebeta-alanine, or salt thereof, in the sustained release formulation isprovided in an amount ranging from about 3.0 to about 7.0 grams.
 27. Thedietary supplement of claim 22 wherein the free beta-alanine, or saltthereof, in the sustained release formulation is provided in an amountranging from about 4.0 to about 6.0 grams.
 28. The dietary supplement ofclaim 22 wherein the sustained release formulation is in a solid, liquidor semi-liquid.
 29. A dietary supplement comprising an amount of amicroencapsulated, sustained release free beta-alanine, or a saltthereof in a microencapsulated, sustained release formulation, whereinfollowing dissolution in 0.1N hydrochloric acid for 1 hour thepercentage of beta-alanine dissolved is less than, or equal to 27%,wherein the free beta-alanine or salt thereof, in the sustained releaseformulation of the dietary supplement is provided in an amount rangingfrom about 0.1 to about 10.0 grams.
 30. The dietary supplement of claim29 wherein the free beta-alanine, or salt thereof, in the sustainedrelease formulation is provided in an amount ranging from about 1.0 toabout 10.0 grams.
 31. The dietary supplement of claim 29 wherein thefree beta-alanine, or salt thereof, in the sustained release formulationis provided in an amount ranging from about 1.0 to about 9.0 grams. 32.The dietary supplement of claim 29 wherein the free beta-alanine, orsalt thereof, in the sustained release formulation is provided in anamount ranging from about 2.0 to about 8.0 grams.
 33. The dietarysupplement of claim 29 wherein the free beta-alanine, or salt thereof,in the sustained release formulation is provided in an amount rangingfrom about 3.0 to about 7.0 grams.
 34. The dietary supplement of claim29 wherein the free beta-alanine, or salt thereof, in the sustainedrelease formulation is provided in an amount ranging from about 4.0 toabout 6.0 grams.
 35. The dietary supplement of claim 29 wherein thesustained release formulation is in a solid, liquid or semi-liquid. 36.A dietary supplement comprising an amount of a microencapsulated,sustained release free beta-alanine, or a salt thereof in amicroencapsulated, sustained release formulation, wherein followingdissolution in 0.1N hydrochloric acid for 2 hours the percentage ofbeta-alanine dissolved is less than, or equal to 47%, wherein the freebeta-alanine or salt thereof, in the sustained release formulation ofthe dietary supplement is provided in an amount ranging from about 0.1to about 10.0 grams.
 37. The dietary supplement of claim 36 wherein thefree beta-alanine, or salt thereof, in the sustained release formulationis provided in an amount ranging from about 1.0 to about 10.0 grams. 38.The dietary supplement of claim 36 wherein the free beta-alanine, orsalt thereof, in the sustained release formulation is provided in anamount ranging from about 1.0 to about 9.0 grams.
 39. The dietarysupplement of claim 36 wherein the free beta-alanine, or salt thereof,in the sustained release formulation is provided in an amount rangingfrom about 2.0 to about 8.0 grams.
 40. The dietary supplement of claim36 wherein the free beta-alanine, or salt thereof, in the sustainedrelease formulation is provided in an amount ranging from about 3.0 toabout 7.0 grams.
 41. The dietary supplement of claim 36 wherein the freebeta-alanine, or salt thereof, in the sustained release formulation isprovided in an amount ranging from about 4.0 to about 6.0 grams.
 42. Thedietary supplement of claim 36 wherein the sustained release formulationis in a solid, liquid or semi-liquid.
 43. A dietary supplementcomprising an amount of a microencapsulated, sustained release freebeta-alanine, or a salt thereof in a microencapsulated, sustainedrelease formulation, wherein following dissolution in 0.1N hydrochloricacid for 3 hours the percentage of beta-alanine dissolved is less than,or equal to 63%, wherein the free beta-alanine or salt thereof, in thesustained release formulation of the dietary supplement is provided inan amount ranging from about 0.1 to about 10.0 grams.
 44. The dietarysupplement of claim 43 wherein the free beta-alanine, or salt thereof,in the sustained release formulation is provided in an amount rangingfrom about 1.0 to about 10.0 grams.
 45. The dietary supplement of claim43 wherein the free beta-alanine, or salt thereof, in the sustainedrelease formulation is provided in an amount ranging from about 1.0 toabout 9.0 grams.
 46. The dietary supplement of claim 43 wherein the freebeta-alanine, or salt thereof, in the sustained release formulation isprovided in an amount ranging from about 2.0 to about 8.0 grams.
 47. Thedietary supplement of claim 43 wherein the free beta-alanine, or saltthereof, in the sustained release formulation is provided in an amountranging from about 3.0 to about 7.0 grams.
 48. The dietary supplement ofclaim 43 wherein the free beta-alanine, or salt thereof, in thesustained release formulation is provided in an amount ranging fromabout 4.0 to about 6.0 grams.
 49. The dietary supplement of claim 43wherein the sustained release formulation is in a solid, liquid orsemi-liquid.
 50. A dietary supplement comprising an amount of amicroencapsulated, sustained release free beta-alanine, or a saltthereof in a microencapsulated, sustained release formulation, whereinfollowing dissolution in 0.1N hydrochloric acid for 6 hours thepercentage of beta-alanine dissolved is less than, or equal to 82%,wherein the free beta-alanine or salt thereof, in the sustained releaseformulation of the dietary supplement is provided in an amount rangingfrom about 0.1 to about 10.0 grams.
 51. The dietary supplement of claim50 wherein the free beta-alanine, or salt thereof, in the sustainedrelease formulation is provided in an amount ranging from about 1.0 toabout 10.0 grams.
 52. The dietary supplement of claim 50 wherein thefree beta-alanine, or salt thereof, in the sustained release formulationis provided in an amount ranging from about 1.0 to about 9.0 grams. 53.The dietary supplement of claim 50 wherein the free beta-alanine, orsalt thereof, in the sustained release formulation is provided in anamount ranging from about 2.0 to about 8.0 grams.
 54. The dietarysupplement of claim 50 wherein the free beta-alanine, or salt thereof,in the sustained release formulation is provided in an amount rangingfrom about 3.0 to about 7.0 grams.
 55. The dietary supplement of claim50 wherein the free beta-alanine, or salt thereof, in the sustainedrelease formulation is provided in an amount ranging from about 4.0 toabout 6.0 grams.
 56. The dietary supplement of claim 50 wherein thesustained release formulation is in a solid, liquid or semi-liquid.